Chemically programmable immunity

ABSTRACT

The present invention is related to methods and compositions that are capable of immediately immunizing a human or animal against any molecule or compound. The present invention comprises an immunity linker molecule with at least two sites; (1) a first binding site that binds to an immune system molecule in a human or animal that has been preimmunized against the first binding site, and (2) one or more second binding sites that bind specifically to a desired compound or molecule. The first binding site and the second binding site(s) are linked by a linker portion of the molecule.

FIELD OF THE INVENTION

The present invention relates to methods and compositions for providingimmediate immunity to any desired antigen. The present invention alsoprovides methods and compositions for treating a wide variety ofdiseases without having to wait for an immune response to be mounted bythe human or animal being exposed to the disease.

BACKGROUND OF THE INVENTION

The term “antigen” is defined as anything that can serve as a target foran immune response. The immune response can be either cellular orhumoral. The term “vaccine” is defined herein as a suspension orsolution of antigenic moieties, usually consisting of infectious agents,or some part of the infectious agents, that is injected into the body toproduce active immunity. The antigenic moiety making up the vaccine canbe either a microorganism or a natural product purified from amicroorganism, a synthetic product or a genetically engineered protein,peptide, polysaccharide or similar product. The term “cell mediatedimmunity” is defined as an immune response mediated by cells rather thanby antibody. It includes, but is not limited to, delayed typehypersensitivity and cytotoxic T cells. The term “adjuvant” as usedherein is any substance whose admixture with an injected immunogenincreases or otherwise modifies the immune response. A “hapten” isdefined antibodies or T cells, but the hapten itself is usually notimmunogenic. Most haptens are small molecules or small parts of largemolecules, but some macromolecules can also function as haptens. Theterm “conjugation” is defined herein as the covalent or other form oflinking two or more molecules. It can be accomplished either by chemicalmeans or in vivo by biologic means such as genetic engineering.

The process of immunization has been used for over a hundred years toprotect humans and animals against disease. The process generallycomprises injecting an antigen that is related to the pathogen in thehuman or animal and waiting an appropriate amount of time, allowing thehuman or animal in which the pathogen was injected to mount an immuneresponse. The time required for mounting an immune response normally isbetween approximately two weeks and several months for most antigens. Inmost cases, a booster administration of the antigen is required tomaintain the immune response. This booster is normally given weeks ormonths after the initial administration of the antigen. Thus,immunization is of little use for immediate treatment of a disease.

A separate immunization procedure must be made for each pathogen,although in some cases several antigens are included in a singlevaccine. Every immunization carries with it a certain amount of riskthat must be considered before any immunization is recommended on awide-scale basis.

What is needed is a method of immunizing a human or animal that canresult in an immediate immune response. In addition, a method ofimmunizing a human or animal by a single immunization would greatlyreduce the inherent risks in the vaccination procedure.

SUMMARY OF THE INVENTION

The present intention provides methods and compositions for theimmediate and specific immunization of a human or animal against apathogen or other undesired substance. The present invention, in oneembodiment, is designated an “immunity linker molecule” and comprises amolecule with multiple sites; a first binding site on the compound thatis antigenic and is capable of mounting an immune response in a human oranimal. After immunization of the human or animal, first binding sitewill then bind specifically to an antibody or other immune molecule thatwas induced by the immunization process. The molecule has a secondbinding site or sites that are capable of binding to one or moredesignated compounds. The present invention also includes a compoundthat contains only the first binding site or immunogenic site that ispresent in the immunity linker molecule. This compound that containsonly the first binding site or antigenic site is designated herein as“the immunizing molecule”.

According to the present invention, the immunity linker molecule can bemade in several ways. The immunizing molecule with the first bindingsite can be physically linked or conjugated to the molecule with thesecond binding sites to the pathogen or other undesired substance. Inanother embodiment, the immunity linker molecule can be produced ormanufactured as a single molecule containing the first binding site orimmunizing site and the second binding sites. The immunity linkermolecule can be any type of compound including protein, nucleic acid ora combination thereof. The first binding sight can be a hapten that isconjugated to a larger molecule.

In practicing the present invention, the human or animal is firstimmunized conventionally against the immunizing molecule. This processincludes administering the molecule to the human or animal and thenwaiting an appropriate amount of time for an immune response to bemounted in the human or animal. If necessary, the immunizing moleculecan be administered with an adjuvant and/or a booster may be given tothe animal at appropriate times. These methods of immunizing a human oranimal are well known to one of ordinary skill in the art. The human oranimal that has been immunized against the immunizing molecule now hasantibodies that will bind the immunizing molecule when it is present inthe blood or other fluid.

When the preimmunized human or animal is challenged with a pathogen ortoxic substance, an immunity linker molecule that contains a bindingsite to the pathogen or toxic substance is administered to the human oranimal. The immunity linker molecule binds at one site to the antibodythat was previously induced, and binds to the pathogen at the secondsite thereby providing an immune complex of the antibody bound to theimmunity linker molecule which is now bound to the pathogen. The bodynow recognizes the immune complexes and processes them in a normalmanner.

Accordingly, it is an object of the present invention to provide amethod and composition for the immediate and specific immunization of ahuman or animal.

It is yet another object of the present invention to provide a methodand composition for immediately immunizing an immunologically naivehuman or animal.

It is another object of the present invention to provide a method andcomposition that enables one to quickly and easily select a desiredantigen and immediately immunize the human or animal against thatantigen.

Another object of the present invention is to provide a method andcomposition that will only require a single immunization to protectagainst a wide variety of pathogens and toxic substances, therebyreducing the risks of multiple vaccinations.

Yet another object of the present invention is to provide a method andcomposition that will allow health care professionals to immediatelyimmunize a patient against a wide variety of pathogens and/or toxins.

These and other objects, features and advantages of the presentinvention will become apparent after a review of the following detaileddescription of the disclosed embodiment.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 illustrates the structure of the immunity linker molecule.

FIG. 2 illustrates the immunity linker molecule bound at one site to anantibody and, at a second site, to a desired molecule, thereby formingan immune complex.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is related to methods and compositions that arecapable of immediately immunizing a human or animal against any moleculeor compound. The present invention comprises an immunity linker moleculewith at least two sites; (1) a first binding site that binds to animmune system molecule in a human or animal that has been preimmunizedagainst the first binding site, and (2) one or more second binding sitesthat bind specifically to a desired compound or molecule. The firstbinding site and the second binding site(s) are linked by a linkerportion of the molecule.

The present invention comprises methods and compositions for theimmediate and specific immunization of a human or animal against apathogen or other undesired substance. According to the presentinvention, a human or animal can be immediately immunized against achosen antigen simply by administering to the human or animal theimmunity linker molecule with the appropriate second binding site.According to the present invention, one can provide immediate immunityto any chosen antigen on the basis of the pre-existing immunity to theimmunizing molecule by administration of a synthetic chemical linker.

In practicing the present invention, the human or animal is firstimmunized conventionally against the immunizing molecule. This processincludes appropriately administering the molecule to the human or animaland then waiting an appropriate amount of time for an immune response tobe mounted in the human or animal. The preferred method of administeringthe immunizing molecule is by injection. If necessary, the immunizingmolecule can be administered with an adjuvant and/or a booster may begiven to the animal at appropriate times. These methods of theimmunizing a human or animal are well known to one of ordinary skill inthe art. The human or animal that has been immunized against theimmunizing molecule now has antibodies that will bind the immunizingmolecule when it is present in the blood or other bodily fluid.

The immunizing molecule optionally can be administered with agents suchas adjuvants, preservatives, diluents, emulsifiers, stabilizers, andother known components that are known and used in immunizationprocedures in the prior art. Any adjuvant system known in the art can beused in the composition of the present invention. Such adjuvantsinclude, but are not limited to, Freund's incomplete adjuvant, Freund'scomplete adjuvant, polydispersed β-(1,4) linked acetylated mannan(“Acemannan”), Titermax® (polyoxyethylene-polyoxypropylene copolymeradjuvants from CytRx Corporation), modified lipid adjuvants from ChironCorporation, saponin derivative adjuvants from Cambridge Biotech, killedBordetella pertussis, the lipopolysaccharide (LPS) of gram-negativebacteria, large polymeric anions such as dextran sulfate, and inorganicgels such as alum, aluminum hydroxide, or aluminum phosphate. Apreferred adjuvant system is Freund's incomplete adjuvant. Anotherpreferred adjuvant system is Freund's complete adjuvant. The method ofimmunization and the adjuvants used are not critical to the invention.Thus, any method known in the art can be used, and any adjuvant systemknown in the art can be used.

According to the present invention, immediate immunity to, for example,a pathogen, can be established in a human or animal that isimmunologically naive to the pathogen by administering to the human oranimal that has been immunized against the immunizing molecule theimmunity linker molecule that contains a binding site to the pathogen.The immunity linker molecule binds at one site to the antibody that waspreviously induced, and binds to the pathogen at the second site,thereby providing an immune complex of the antibody bound to theimmunity linker molecule which is now bound to the pathogen. The bodynow recognizes the immune complexes and processes the complexes in anormal manner.

According to the present invention, the immunity linker molecule can bemade in several ways. The immunizing molecule can be physically linkedor conjugated to the molecule with the binding sites to the desiredsubstance. In another embodiment, the immunity linker molecule can beproduced or manufactured as a single molecule containing the multiplesites. In yet another embodiment, the immunity linker molecule consistsof two active ends connected together by a rigid or flexible spacer suchas a double helical region of RNA. The purpose of the spacer is to holdthe two ends of the linker together, while preventing them frominteracting.

The immunity linker molecule can be a protein, peptide, or nucleic acidmolecule or any combination thereof, including, but not limited to, RNAmolecules, DNA molecules or derivatives thereof. Preferably, theimmunity linker molecule is comprised of RNA molecules and are producedaccording to the SELEX process. This process is described completely inthe list of references attached hereto and are included herein byreference in their entirety.

The immunity linker molecule is shown schematically in FIG. 1. Theimmunity linker molecule 10 comprises a first binding site 15 which isantigenic, a linking portion of the molecule 20 and a second bindingsite 35 that is capable of binding a specific molecule. The secondbinding site 35 site or sites are preferably aptamers that have beenproduced by the SELEX process. However, it is to be understood that thesecond binding site does not have to be an aptamer, but can be any typeof molecule that has the desired physical attributes, i.e., the secondbinding site being capable of binding to a specific molecule. It is tobe understood that the immunity linker molecule can have more than onebinding site to a single substance or can have multiple binding sitesagainst multiple substances. The linking portion of the molecule linksthe first binding site 15 and the second binding site 35. The linkingportion 15 of the molecule can be double stranded nucleic acid, butother linking molecules can be used in the present invention. FIG. 2schematically shows the immunity linker molecule with an antibody 40bound to the first binding site 15 of the molecule and a molecule 45bound to the second binding site 35 on the immunity linker molecule 10.

It is to be understood that the immunity linker molecule can be any typeof molecule that is capable of being manipulated so that it is capableof (1) mounting an immunity response, and (2) binding a desired moleculeor molecules. The preferred type of compound is nucleic acid or,preferably, modified nucleic acid such as 2′-fluoro- or2′-amino-2′-deoxypyrimidine containing nucleic acids. Nucleic acidsusing these bases are much more stable than naturally occurring nucleicacids. (See Aptamers as tools in molecular biology and immunology, M.Famulok and G. Mayer, Cur. Top. Micro. Immunobiol., 1999, 243, 123-146.)

The immunity linker molecule can be administered to a patientintramuscularly, subcutaneously, orally, intravenously, or through themucosal membranes. The immunity linker molecule can be use in immunizinga human or animal against a wide variety of substrates, including, butnot limited to, bacteria, fungi, viruses, toxic substances, and drugs.

The present invention is particularly useful in the military wheretroops may be unexpectedly exposed to a pathogen, toxin, or to a toxicchemical substance. Military personnel are preimmunized against theimmunizing molecule, i.e., that portion of the immunity linker moleculethat binds to the antibody. Then, if the military personnel areunexpectedly challenged with a pathogen, the appropriate immunity linkermolecule can be administered to the military personnel, therebyimmediately protecting them against the pathogen or other toxicsubstance. The present invention can be used to prevent and/or treatorganisms including, but not limited to, anthrax, dengue virus, orMarburg virus.

Likewise, pharmacies can have a library of different immunity linkermolecules available for a variety of different pathogens and toxicsubstances. If the patient has been preimmunized against the immunizingportion of the linker, then he or she will be immediately immunizedagainst the pathogen or toxic substances.

It should be understood, of course, that the foregoing relates only topreferred embodiments of the present invention and that numerousmodifications or alterations may be made therein without departing fromthe spirit and the scope of the invention as set forth in thisdisclosure.

1. An immunity linker molecule, the immunity linker molecule containingat least one first site that binds to an immune system molecule and atlease one second site that binds to a desired compound or molecule. 2.The immunity linker molecule of claim 1 wherein the first site binds toan antibody.
 3. The antibody of claim 2 wherein the antibody is from ahuman or animal that has been preimmunized against the first site. 4.The immunity linker molecule of claim 1, wherein the desired compound ormolecule is a microorganism.
 5. The immunity linker molecule of claim 4wherein the microorganism is a bacteria, virus, or fungus.
 6. Theimmunity linker molecule of claim 1, wherein the compound or molecule isa drug.
 7. The immunity linker molecule of claim 1, wherein the immunitylinker molecule is an aptomer.
 8. A method of immunizing a human oranimal against a molecule or compound comprising administering to thehuman or animal an immunity linker molecule, the immunity linkermolecule containing at least one first site that binds to an immunesystem molecule and at lease one second site that binds to a desiredcompound or molecule.
 9. The method of claim 8, wherein the human oranimal has been preimmunized against the first site on the immunitylinker molecule.
 10. The method of claim 8, wherein the immunity thedesired compound or molecule is a microorganism.
 11. The method of claim10 wherein the microorganism is a bacteria, virus, or fungus.
 12. Themethod of claim 8, wherein the compound or molecule is a drug.
 13. Themethod of claim 1, wherein the immunity linker molecule is an aptomer.14. A method of preimmunizing a human or animal comprising immunizing ahuman or animal against a molecule that contains a first site that isimmunogenic, the first site being present on an immune linker molecule,the immune linker molecule also containing at lease one second site thatbinds to a desired compound or molecule.